摘要
根据已报道的人参HMGR(3-羟基-3-甲基戊二酰CoA还原酶)基因cDNA序列设计引物,利用cDNA末端快速扩增(Rapid amplification of cDNA ends,RACE)技术克隆刺五加HMGR基因的全长cDNA序列。运用生物信息学方法对该基因进行分析。通过RT-PCR法检测HMGR在刺五加不同生长发育时期和不同器官中的表达情况。结果克隆到长度为2 217 bp的刺五加HMGR基因cDNA序列,开放阅读框全长1 713 bp,编码570个氨基酸残基,包含HMGR家族的特异性识别序列。HMGR蛋白存在2个跨膜区域。RT-PCR结果显示,刺五加HMGR基因在各生长发育时期和器官中均有表达,但表达量具有显著差异,其中萌芽期的表达量最高,盛花期最低;各器官中,幼茎中表达量最高,是根中的1.58倍。
In order to clone 3-hydroxy-3-methylglutaryl coenzyme A reductase（HMGR） in Eleutherococcus senticosus,the primers were designed according to the reported Panax ginseng HMGR.The full length cDNA was cloned by rapid amplification of cDNA ends（RACE）,and analyzed by bioinformatics.The expression of HMGR in different growth periods and organs of Eleutherococcus senticosus was detected by RT-PCR.E.senticosus HMGR was cloned successfully,with full length of 2 217 bp,containing an ORF spaning 1 713 bp that encoded 570 amino acids.The deduced protein exhibited specific recognition sequence of HMGR family and had two transmembrane regions.RT-PCR results showed that HMGR gene was expressed in various growth periods and organs of E.senticosus,and the expression differed significantly.The highest expression of HMGR was at germination stage and the lowest at full-bloom stage.In organs,the greatest expression was in young stem,1.58 times as much as the lowest in root.
出处
《江苏农业学报》
CSCD
北大核心
2012年第6期1258-1262,共5页
Jiangsu Journal of Agricultural Sciences
基金
国家自然科学基金项目(30701086)
河北省自然科学基金项目(C2009001252)
河北省自然科学基金-石药集团医药联合研究基金项目(H2012401006)
